2-{(R)-2-methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamide crystalline form 1

ABSTRACT

2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamide Crystalline Form 1, ways to make it, compositions comprising it and made using it, and methods of treating patients having disease using it are disclosed.

This application claims priority from U.S. Provisional PatentApplication Ser. No. 60/979,643, filed Oct. 12, 2007, incorporatedherein by reference.

FIELD OF THE INVENTION

This invention pertains to2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamide, alsoreferred to herein as ABT-888, Crystalline Form 1, ways to make it,compositions comprising it and made using it, and methods of treatingpatients having disease using it.

BACKGROUND OF THE INVENTION

Poly(ADP-ribose)polymerase (PARP) has an essential role in facilitatingDNA repair, controlling RNA transcription, mediating cell death andregulating immune response. PARP inhibitors have demonstrated efficacyin a number of disease models, such as, for example,2-((R)-2-methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamide (ABT-888)that has shown to limit tumor growth in tumor-bearing animals.

Crystallinity of ABT-888 may effect, among other physical and mechanicalproperties, its stability, solubility, dissolution rate, hardness,compressability and melting point. Because ease of manufacture andformulation of ABT-888 is dependent on some, if not all, of theseproperties, there is an existing need in the chemical and therapeuticarts for identification of crystalline forms of ABT-888 and ways toreproducibly make them.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows a theoretical powder X-ray diffraction (PXRD) spectrum ofABT-888 Crystalline Form 1.

FIG. 2 shows an experimental PXRD spectrum of ABT-888 Crystalline Form1.

FIG. 3 shows a differential scanning calorimetric (DSC) spectrograph ofABT-888 Crystalline Form 1.

FIG. 4 shows the infra red (FT/IR) spectrum of ABT-888 Crystalline Form1.

FIG. 5 shows the Raman spectrum of ABT-888 Crystalline Form 1.

FIG. 6 shows a theoretical PXRD spectrum of ABT-888 Crystalline Form 2.

FIG. 7 shows an experimental PXRD spectrum of ABT-888 Crystalline Form2.

FIG. 8 shows the infra red (FT/IR) spectrum of ABT-888 Crystalline Form2.

FIG. 9 shows the Raman spectrum of ABT-888 Crystalline Form 2.

SUMMARY OF THE INVENTION

One embodiment of this invention pertains to2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1 which, when measured at about −100° C. in thetetragonal crystal system and P4₃2₁2 space group with radiation at0.7107 Å, is characterized by respective lattice parameter values a, band c of 8.218 Å±0.002 Å, 8.218 Å±0.002 Å and 36.06 Å±0.01 Å and α, βand γ values of 90°.

Another embodiment pertains to2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1 which, when measured at about 25° C. with radiationat 1.54178 Å, is characterized by a powder diffraction pattern havingrespective 2θ values of about 9.9°, 11.0° and 11.8° and one or more thanone additional peaks having respective 2θ values of about 14.6°, 15.2°,18.2°, 19.6°, 20.3°, 21.3°, 22.5°, 22.8°, 24.7°, 28.5° and 29.1°.

Still another embodiment pertains to compositions comprising anexcipient and2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1 which, when measured at about −100° C. in thetetragonal crystal system and P4₃2₁2 space group with radiation at0.7107 Å, is characterized by respective lattice parameter values a, band c of 8.218 Å±0.002 Å, 8.218 Å±0.002 Å and 36.06 Å±0.01 Å and α, βand γ values of 90°.

Another embodiment pertains to compositions comprising an excipient and2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1 which, when measured at about 25° C. with radiationat 1.54178 Å, is characterized by a powder diffraction pattern havingrespective 2θ values of about 9.9°, 11.0° and 11.8° and one or more thanone additional peaks having respective 2θ values of about 14.6°, 15.2°,18.2°, 19.6°, 20.3°, 21.3°, 22.5°, 22.8°, 24.7°, 28.5° and 29.1°.

Still another embodiment pertains to methods of treating cancer in amammal comprising administering thereto, with or without one or morethan one additional anticancer drugs, a therapeutically effective amountof 2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1 which, when measured at about −100° C. in thetetragonal crystal system and P4₃2₁2 space group with radiation at0.7107 Å, is characterized by respective lattice parameter values a,band c of 8.218 Å±0.002 Å, 8.218 Å±0.002 Å and 36.06 Å±0.01 Å and α, βand γ values of 90°.

Still another embodiment pertains to methods of treating cancer in amammal comprising administering thereto, with or without one or morethan one additional anticancer drugs, a therapeutically effective amountof 2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1 which, when measured at about 25° C. with radiationat 1.54178 Å, is characterized by a powder diffraction pattern havingrespective 2θ values of about 9.9°, 11.0° and 11.8° and one or more thanone additional peaks having respective 2θ values of about 14.6°, 15.2°,18.2°, 19.6°, 20.3°, 21.3°, 22.5°, 22.8°, 24.7°, 28.5° and 29.1°.

Still another embodiment pertains to a process for making2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1, comprising:

making 2-((R)-2-methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamide;

providing a mixture comprising2-((R)-2-methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamide andsolvent, wherein the2-((R)-2-methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamide iscompletely dissolved in the solvent;

causing 2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1 to exist in the mixture, which2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1, when isolated and measured at about −100° C. in thetetragonal crystal system and P4₃2₁2 space group with radiation at0.7107 Å, is characterized by respective lattice parameter values a, band c of 8.218 Å±0.002 Å, 8.218 Å±0.002 Å and 36.06 Å±0.01 Å and α, βand γ values of 90°; and

isolating the2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1.

Still another embodiment comprises2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1 prepared by the process of the preceeding embodiment.

Still another embodiment pertains to a process for making2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1, comprising:

making 2-((R)-2-methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamide;

providing a mixture comprising2-((R)-2-methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamide and2-propanol at about 82° C., wherein the2-((R)-2-methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamide iscompletely dissolved in the 2-propanol;

causing 2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1 to exist in the mixture by lowering its temperature,wherein the2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1, when isolated and measured at about −100° C. in thetetragonal crystal system and P4₃2₁2 space group with radiation at0.7107 Å, is characterized by respective lattice parameter values a, band c of 8.218 Å±0.002 Å, 8.218 Å±0.002 Å and α, β and γ values of 90°;and

isolating the2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1.

Still another embodiment comprises2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1 prepared by the process of the preceeding embodiment.

In a process for making2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1 comprising reacting an acid or diacid salt of2-((R)-2-methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamide and abase and crystallization or recrystallization of2-((R)-2-methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamide to2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1, still another embodiment of this invention comprisescrystallizing or recrystallizing2-((R)-2-methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1 from a solid, semisolid, wax or oil form of2-((R)-2-methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamide that ismixed with one or more than one solvent from the deprotonation reaction.

Still another embodiment comprises2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1 prepared by the process of the preceeding embodiment.

In a process for making2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1 comprising reacting the dihydrochloride salt of2-((R)-2-methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamide andsodium bicarbonate in water and n-butanol and subsequently crystallizing2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1 from 2-propanol, still another embodiment of thisinvention comprises crystallizing2-((R)-2-methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1 from a solid, semisolid, wax or oil form of2-((R)-2-methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamide that ismixed with water or n-butanol.

Still another embodiment comprises2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1 prepared by the process of the preceeding embodiment.

Still another embodiment comprises a mono-acid or di-acid salt of2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamide for usein preparing2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1.

Still another embodiment comprises2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamidedihydrochloride for use in preparing2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1.

Still another embodiment comprises amorphous2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamide for usein preparing2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1.

Still another embodiment comprises2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 2 for use in preparing2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1.

DETAILED DESCRIPTION OF THE INVENTION

This invention pertains to discovery of2-((R)-2-methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamide, alsoreferred to herein as ABT-888, Crystalline Form 1, ways to make it, waysto characterize it, compositions containing it and made with it, andmethods of treating cancer using it. The terms“2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamide” and“ABT-888” are meant to be used interchangeably.

The terms “ABT-888” and “an ABT-888” without any indicia ofcrystallinity or non-crystallinity associated with it, as used herein,mean amorphous ABT-888, a crystalline ABT-888 such as ABT-888Crystalline Form 1 or ABT-888 Crystalline Form 2, microcrystallineABT-888, ABT-888 in solution, a semisolid, wax or oil form of ABT-888,mixtures thereof and the like.

The terms “crystalline” and “microcrystalline,” as used herein, meanhaving a regularly repeating arrangement of molecules which ismaintained over a long range or external face planes.

The term “crystalline ABT-888,” as used herein, means a particularcrystalline ABT-888, including the crystalline ABT-888 of thisinvention, i.e. ABT-888 Crystalline Form 1.

The term “crystalline ABT-888 of this invention,” as used herein, meansthe most thermodynamically stable crystalline form of ABT-888 at 25° C.,i.e. ABT-888 Crystalline Form 1 which, when measured at about −100° C.in the tetragonal crystal system and P4₃2₁2 space group with radiationat 0.7107 Å, is characterized by respective lattice parameter values a,b and c of 8.218 Å±0.002 Å, 8.218 Å±0.002 Å and 36.06 Å±0.01 Å and α, βand γ values of 90° or which, when measured at about 25° C. withradiation at 1.54178 Å, is characterized by a powder diffraction patternhaving respective 2θ values of about 9.9°, 11.0° (1,0,1) and 11.8°(1,0,2) and one or more than one additional peaks having respective 2θvalues of about 14.6° (1,0,4), 15.2° (1,1,0), 18.2° (1,1,4), 19.6°(1,1,5), 20.3° (1,0,7), 21.3° (1,1,6), 22.5° (1,0,8), 22.8° (2,0,3),24.7° (1,0,9), 28.5° (2,1,6) and 29.1° (1,1,10), wherein mostabsorptions are shown with their corresponding Miler Index (h,k,l)values.

The term “amorphous,” as used herein, means a supercooled liquidsubstance or a viscous liquid which appears as a solid but does not havea regularly repeating arrangement of molecules which is maintained overa long range. Amorphous substances do not have melting points but rathersoften or flow above a certain temperature known as the glass transitiontemperature.

The term “semisolid ABT-888,” as used herein, means a combination ofABT-888 and solvent in a gelatinous enough state to prevent passagethrough a semi-permeable membrane or filter.

It is meant to be understood that when peak positions are further usedto identify a particular crystalline form of a compound when unit cellparameters of the compound are used in combination therewith, any onepeak position or combination of peak positions may be used to furtheridentify the particular crystalline form.

Unless stated otherwise, percentages herein are weight/weight (w/w)percentages.

The term “solvent,” as used herein, means a liquid in which a compoundis soluble or partially soluble enough at a given concentration todissolve or partially dissolve the compound.

The term “anti-solvent,” as used herein, means a liquid in which acompound is insoluble enough at a given concentration to be effectivefor precipitating that compound from a solution.

Solvents and anti-solvents may be mixed with or without separation ofphases.

It is meant to be understood that, because many solvents andanti-solvents contain impurities, the level of impurities in solventsand anti-solvents for the practice of this invention, if present, are ata low enough concentration that they do not interfere with the intendeduse of the solvent in which they are present.

The term “acid,” as used herein, means a compound having at least oneacidic proton. Examples of acids for the practice of this inventioninclude, but are not limited to, hydrochloric acid, hydrobromic acid,trifluoroacetic acid, trichloroacetic acid, sulfuric acid, phosphoricacid and the like.

The term “base,” as used herein, means a compound capable of accepting aproton. Examples of bases for the practice of this invention include,but are not limited to, sodium carbonate, sodium bicarbonate, potassiumcarbonate, potassium bicarbonate triethylamine, diisopropylethylamineand the like.

Causing ABT-888 Crystalline Form 1 to exist in a mixture comprisingABT-888 and solvent, wherein the ABT-888 has completely dissolved, isknown as nucleation.

For the practice of this invention, nucleation may be made to occur bymeans such as solvent removal, temperature change, solvent-miscibleanti-solvent addition, solvent-immiscible anti-solvent addition, seedcrystal addition of ABT-888 Crystalline Form 1, chafing or scratchingthe interior of the container, preferably a glass container, in whichnucleation is meant to occur with an implement such as a glass rod or aglass bead or beads, or a combination of the foregoing.

For the practice of this invention, nucleation may be followed bycrystal growth, accompanied by crystal growth, or followed andaccompanied by crystal growth during which, and as a result of which,the percentage of ABT-888 Crystalline Form 1 increases.

It is meant to be understood that airborne seeds (seed crystals) ofcrystalline ABT-888 Crystalline Form 1 may also cause nucleation in amixture of ABT-888 Crystalline Form 1 and solvent in which the ABT-888his completely dissolved.

The term “seed crystal,” as used herein, means a particular crystallineform of a substance having mass. It is meant to be understood that sucha crystal may be small enough to be airborne or invisible to the eyewithout means of detection.

The term “isolating” as used herein, means separating ABT-888Crystalline Form 1 and solvent, anti-solvent, or a mixture of solventanti-solvent. This is typically accomplished by means such ascentrifugation, filtration with or without vacuum, filtration withpositive pressure, distillation, evaporation or a combination thereof.

Therapeutically acceptable amounts of ABT-888 Crystalline Form 1 dependon recipient of treatment, disorder being treated and severity thereof,composition containing it, time of administration, route ofadministration, duration of treatment, its potency, its rate ofclearance and whether or not another drug is co-administered. The amountof ABT-888 Crystalline Form 1 used to make a composition to beadministered daily to a patient in a single dose or in divided doses isfrom about 0.03 to about 200 mg/kg body weight. Single dose compositionscontain these amounts or a combination of submultiples thereof.

ABT-888 Crystalline Form 1 may be administered with or without anexcipient. Excipients include but are not limited to, for example,encapsulating materials and additives such as absorption accelerators,antioxidants, binders, buffers, coating agents, coloring agents,diluents, disintegrating agents, emulsifiers, extenders, fillers,flavoring agents, humectants, lubricants, perfumes, preservatives,propellants, releasing agents, sterilizing agents, sweeteners,solubilizers, wetting agents, mixtures thereof and the like.

Excipients for preparation of compositions comprising or made withABT-888 Crystalline Form 1 to be administered orally in solid dosageform include, for example, agar, alginic acid, aluminum hydroxide,benzyl alcohol, benzyl benzoate, 1,3-butylene glycol, carbomers, castoroil, cellulose, cellulose acetate, cocoa butter, corn starch, corn oil,cottonseed oil, cross-povidone, diglycerides, ethanol, ethyl cellulose,ethyl laureate, ethyl oleate, fatty acid esters, gelatin, germ oil,glucose, glycerol, groundnut oil, hydroxypropylmethyl cellulose,isopropanol, isotonic saline, lactose, magnesium hydroxide, magnesiumstearate, malt, mannitol, monoglycerides, olive oil, peanut oil,potassium phosphate salts, potato starch, povidone, propylene glycol,Ringer's solution, safflower oil, sesame oil, sodium carboxymethylcellulose, sodium phosphate salts, sodium lauryl sulfate, sodiumsorbitol, soybean oil, stearic acids, stearyl fumarate, sucrose,surfactants, talc, tragacanth, tetrahydrofurfuryl alcohol,triglycerides, water, mixtures thereof and the like.

Excipients for preparation of compositions comprising or made withABT-888 Crystalline Form 1 to be administered ophthalmically or orallyin liquid dosage forms include, for example, 1,3-butylene glycol, castoroil, corn oil, cottonseed oil, ethanol, fatty acid esters of sorbitan,germ oil, groundnut oil, glycerol, isopropanol, olive oil, polyethyleneglycols, propylene glycol, sesame oil, water, mixtures thereof and thelike.

Excipients for preparation of compositions comprising or made withABT-888 Crystalline Form 1 to be administered osmotically include, forexample, chlorofluorohydrocarbons, ethanol, water, mixtures thereof andthe like.

Excipients for preparation of compositions comprising or made withABT-888 Crystalline Form 1 to be administered parenterally include, forexample, 1,3-butanediol, castor oil, corn oil, cottonseed oil, dextrose,germ oil, groundnut oil, liposomes, oleic acid, olive oil, peanut oil,Ringer's solution, safflower oil, sesame oil, soybean oil, U.S.P. orisotonic sodium chloride solution, water, mixtures thereof and the like.

Excipients for preparation of compositions comprising or made withABT-888 Crystalline Form 1 to be administered rectally or vaginallyinclude, but are not limited to, cocoa butter, polyethylene glycol, wax,mixtures thereof and the like.

Preparation of ABT-888 and its utility as a PARP inhibitor is describedin commonly-owned United States Patent Application Publication No.2006/0229289.

The hydrochloride salt of ABT-888 is disclosed in commonly-owned U.S.Patent Application Publication No. 2006/0229289.

ABT-888 Crystalline Form 1 is useful when administered with anticancerdrugs such as alkylating agents, angiogenesis inhibitors, antibodies,antimetabolites, antimitotics, antiproliferatives, aurora kinaseinhibitors, Bcr-Abl kinase inhibitors, biologic response modifiers,cyclin-dependent kinase inhibitors, cell cycle inhibitors,cyclooxygenase-2 inhibitors, leukemia viral oncogene homolog (ErbB2)receptor inhibitors, growth factor inhibitors, heat shock protein(HSP)-90 inhibitors, histone deacetylase (HDAC) inhibitors inhibitors,hormonal therapies, immunologicals, intercalating antibiotics, kinaseinhibitors, mammalian target of rapomycin inhibitors, mitogen-activatedextracellular signal-regulated kinase inhibitors, non-steroidalanti-inflammatory drugs (NSAID's), platinum chemotherapeutics, polo-likekinase inhibitors, proteasome inhibitors, purine analogs, pyrimidineanalogs, receptor tyrosine kinase inhibitors, retinoids/deltoids plantalkaloids, topoisomerase inhibitors and the like.

Alkylating agents include altretamine, AMD-473, AP-5280, apaziquone,bendamustine, brostallicin, busulfan, carboquone, carmustine (BCNU),chlorambucil, Cloretazine™ (VNP 40101M), cyclophosphamide, decarbazine,estramustine, fotemustine, glufosfamide, ifosfamide, KW-2170, lomustine(CCNU), mafosfamide, melphalan, mitobronitol, mitolactol, nimustine,nitrogen mustard N-oxide, ranimustine, temozolomide, thiotepa,treosulfan, trofosfamide and the like.

Angiogenesis inhibitors include endothelial-specific receptor tyrosinekinase (Tie-2) inhibitors, epidermal growth factor receptor (EGFR)inhibitors, insulin growth factor-2 receptor (IGFR-2) inhibitors, matrixmetalloproteinase-2 (MMP-2) inhibitors, matrix metalloproteinase-9(MMP-9) inhibitors, platelet-derived growth factor receptor (PDGFR)inhibitors, thrombospondin analogs vascular endothelial growth factorreceptor tyrosine kinase (VEGFR) inhibitors and the like.

Aurora kinase inhibitors include AZD-1152, MLN-8054, VX-680 and thelike.

Bcr-Abl kinase inhibitors include DASATINIB® (BMS-354825), GLEEVEC®(imatinib) and the like.

CDK inhibitors include AZD-5438, BMI-1040, BMS-032, BMS-387, CVT-2584,flavopyridol, GPC-286199, MCS-5A, PD0332991, PHA-690509, seliciclib(CYC-202, R-roscovitine), ZK-304709 and the like.

COX-2 inhibitors include ABT-963, ARCOXIA® (etoricoxib), BEXTRA®(valdecoxib), BMS347070, CELEBREX™ (celecoxib), COX-189 (lumiracoxib),CT-3, DERAMAXX® (deracoxib), JTE-522,4-methyl-2-(3,4-dimethylphenyl)-1-(4-sulfamoylphenyl-1H-pyrrole), MK-663(etoricoxib), NS-398, parecoxib, RS-57067, SC-58125, SD-8381, SVT-2016,S-2474, T-614, VIOXX® (rofecoxib) and the like.

EGFR inhibitors include ABX-EGF, anti-EGFr immunoliposomes, EGF-vaccine,EMD-7200, ERBITUX® (cetuximab), HR3, IgA antibodies, IRESSA®(gefitinib), TARCEVA® (erlotinib or OSI-774), TP-38, EGFR fusionprotein, TYKERB® (lapatinib) and the like.

ErbB2 receptor inhibitors include CP-724-714, CI-1033 (canertinib),Herceptin® (trastuzumab), TYKERB® (lapatinib), OMNITARG® (2C4,petuzumab), TAK-165, GW-572016 (ionafamib), GW-282974, EKB-569, PI-166,dHER2 (HER2 vaccine), APC-8024 (HER-2 vaccine), anti-HER/2neu bispecificantibody, B7.her2IgG3, AS HER2 trifunctional bispecfic antibodies, mABAR-209, mAB 2B-1 and the like.

Histone deacetylase inhibitors include depsipeptide, LAQ-824, MS-275,trapoxin, suberoylanilide hydroxamic acid (SAHA), TSA, valproic acid andthe like.

HSP-90 inhibitors include 17-AAG-nab, 17-AAG, CNF-101, CNF-1010,CNF-2024, 17-DMAG, geldanamycin, IPI-504, KOS-953, MYCOGRAB®,NCS-683664, PU24FCl, PU-3, radicicol, SNX-2112, STA-9090 VER49009 andthe like.

MEK inhibitors include ARRY-142886, ARRY-438162 PD-325901, PD-98059 andthe like.

mTOR inhibitors include AP-23573, CCI-779, everolimus, RAD-001,rapamycin, temsirolimus and the like.

Non-steroidal anti-inflammatory drugs include AMIGESIC® (salsalate),DOLOBID® (diflunisal), MOTRIN® (ibuprofen), ORUDIS® (ketoprofen),RELAFEN® (nabumetone), FELDENE® (piroxicam) ibuprofin cream, ALEVE® andNAPROSYN® (naproxen), VOLTAREN® (diclofenac), INDOCIN® (indomethacin),CLINORIL® (sulindac), TOLECTIN® (tolmetin), LODINE® (etodolac), TORADOL®(ketorolac), DAYPRO® (oxaprozin) and the like.

PDGFR inhibitors include C-451, CP-673, CP-868596 and the like.

Platinum chemotherapeutics include cisplatin, ELOXATIN® (oxaliplatin)eptaplatin, lobaplatin, nedaplatin, PARAPLATIN® (carboplatin),satraplatin and the like.

Polo-like kinase inhibitors include BI-2536 and the like.

Thrombospondin analogs include ABT-510, ABT-567, ABT-898, TSP-1 and thelike.

VEGFR inhibitors include AVASTIN® (bevacizumab), ABT-869, AEE-788,ANGIOZYME™, axitinib (AG-13736), AZD-2171, CP-547,632, IM-862, Macugen(pegaptamib), NEXAVAR® (sorafenib, BAY43-9006), pazopanib (GW-786034),(PTK-787, ZK-222584), SUTENT® (sunitinib, SU-11248), VEGF trap,vatalanib, ZACTIMA™ (vandetanib, ZD-6474) and the like.

Antimetabolites include ALIMTA® (premetrexed disodium, LY231514, MTA),5-azacitidine, XELODA® (capecitabine), carmofur, LEUSTAT® (cladribine),clofarabine, cytarabine, cytarabine ocfosfate, cytosine arabinoside,decitabine, deferoxamine, doxifluridine, eflornithine, EICAR,enocitabine, ethnylcytidine, fludarabine, hydroxyurea, 5-fluorouracil(5-FU) alone or in combination with leucovorin, GEMZAR® (gemcitabine),hydroxyurea, ALKERAN® (melphalan), mercaptopurine, 6-mercaptopurineriboside, methotrexate, mycophenolic acid, nelarabine, nolatrexed,ocfosate, pelitrexol, pentostatin, raltitrexed, Ribavirin, triapine,trimetrexate, S-1, tiazofurin, tegafur, TS-1, vidarabine, UFT and thelike.

Antibiotics include intercalating antibiotics aclarubicin, actinomycinD, amrubicin, annamycin, adriamycin, BLENOXANE® (bleomycin),daunorubicin, CAELYX® or MYOCET® (doxorubicin), elsamitrucin, epirbucin,glarbuicin, ZAVEDOS® (idarubicin), mitomycin C, nemorubicin,neocarzinostatin, peplomycin, pirarubicin, rebeccamycin, stimalamer,streptozocin, VALSTAR® (valrubicin), zinostatin and the like.

Topoisomerase inhibitors include aclarubicin, 9-aminocamptothecin,amonafide, amsacrine, becatecarin, belotecan, BN-80915, CAMPTOSAR®(irinotecan hydrochloride), camptothecin, CARDIOXANE® (dexrazoxine),diflomotecan, edotecarin, ELLENCE® or PHARMORUBICIN® (epirubicin),etoposide, exatecan, 10-hydroxycamptothecin, gimatecan, lurtotecan,mitoxantrone, orathecin, pirarbucin, pixantrone, rubitecan, sobuzoxane,SN-38, tafluposide, topotecan and the like.

Antibodies include AVASTIN® (bevacizumab), CD40-specific antibodies,chTNT-1/B, denosumab, ERBITUX® (cetuximab), HUMAX-CD4® (zanolimumab),IGF1R-specific antibodies, lintuzumab, PANOREX® (edrecolomab), RENCAREX®(WX G250), RITUXAN® (rituximab), ticilimumab, trastuzimab and and thelike.

Hormonal therapies include ARIMIDEX® (anastrozole), AROMASIN®(exemestane), arzoxifene, CASODEX® (bicalutamide), CETROTIDE®(cetrorelix), degarelix, deslorelin, DESOPAN® (trilostane),dexamethasone, DROGENIL®, (flutamide), EVISTA® (raloxifene), fadrozole,FARESTON® (toremifene), FASLODEX® (fulvestrant),FMARA®, (letrozole),formestane, glucocorticoids, HECTOROL® or RENAGEL® (doxercalciferol),lasofoxifene, leuprolide acetate, MEGACE® (megesterol), MIFEPREX®(mifepristone), NILANDRON™ (nilutamide), NOLVADEX® (tamoxifen citrate),PLENAXIS™ (abarelix), predisone, PROPECIA® (finasteride), rilostane,SUPREFACT® (buserelin), TRELSTAR® (luteinizing hormone releasing hormone(LHRH)), vantas, VETORYL®, (trilostane or modrastane), ZOLADEX®(fosrelin, goserelin) and the like.

Deltoids and retinoids include seocalcitol (EB1089, CB1093),lexacalcitrol (KH1060), fenretinide, PANRETIN® (aliretinoin), ATRAGEN®(liposomal tretinoin), TARGRETIN® (bexarotene), LGD-1550 and the like.

Plant alkaloids include, but are not limited to, vincristine,vinblastine, vindesine, vinorelbine and the like.

Proteasome inhibitors include VELCADE® (bortezomib), MG132, NPI-0052,PR-171 and the like.

Examples of immunologicals include interferons and otherimmune-enhancing agents. Interferons include interferon alpha,interferon alpha-2a, interferon alpha-2b, interferon beta, interferongamma-1a, ACTIMMUNE® (interferon gamma-1b), or interferon gamma-n1,combinations thereof and the like. Other agents include ALFAFERONE®,BAM-002, BEROMUN® (tasonermin), BEXXAR® (tositumomab), CamPath®(alemtuzumab), CTLA4 (cytotoxic lymphocyte antigen 4), decarbazine,denileukin, epratuzumab, GRANOCYTE® (lenograstim), lentinan, leukocytealpha interferon, imiquimod, MDX-010, melanoma vaccine, mitumomab,molgramostim, MYLOTARG™ (gemtuzumab ozogamicin), NEUPOGEN® (filgrastim),OncoVAC-CL, OvaRex® (oregovomab), pemtumomab (Y-muHMFG1), PROVENGE®,sargaramostim, sizofilan, teceleukin, TheraCys®, ubenimex, VIRULIZIN®,Z-100, WF-10, PROLEUKIN® (aldesleukin), ZADAXIN® (thymalfasin), ZENAPAX®(daclizumab), ZEVALIN® (90Y-Ibritumomab tiuxetan) and the like.

Biological response modifiers are agents that modify defense mechanismsof living organisms or biological responses, such as survival, growth,or differentiation of tissue cells to direct them to have anti-tumoractivity and include include krestin, lentinan, sizofiran, picibanilPF-3512676 (CpG-8954), ubenimex and the like.

Pyrimidine analogs include cytarabine (ara C or Arabinoside C), cytosinearabinoside, doxifluridine, FLUDARA® (fludarabine), 5-FU(5-fluorouracil), floxuridine, GEMZAR® (gemcitabine), TOMUDEX®(ratitrexed), TROXATYL™ (triacetyluridine troxacitabine) and the like.

Purine analogs include LANVIS® (thioguanine) and PURI-NETHOL®(mercaptopurine).

Antimitotic agents include batabulin, epothilone D (KOS-862),N-(2-((4-hydroxyphenyl)amino)pyridin-3-yl)-4-methoxybenzenesulfonamide,ixabepilone (BMS 247550), paclitaxel, TAXOTERE® (docetaxel), PNU100940(109881), patupilone, XRP-9881, vinflunine, ZK-EPO and the like.

Compounds of the present invention are also intended to be used as aradiosensitizer that enhances the efficacy of radiotherapy. Examples ofradiotherapy include, but are not limited to, external beamradiotherapy, teletherapy, brachtherapy and sealed and unsealed sourceradiotherapy.

Additionally, ABT-888 Crystalline Form 1 may be combined with otherchemptherapeutic agents such as ABRAXANE™ (ABI-007), ABT-100 (farnesyltransferase inhibitor), ADVEXIN®, ALTOCOR® or MEVACOR® (lovastatin),AMPLIGEN® (poly I:poly C12U, a synthetic RNA), APTOSYN™ (exisulind),AREDIA® (pamidronic acid), arglabin, L-asparaginase, atamestane(1-methyl-3,17-dione-androsta-1,4-diene), AVAGE® (tazarotne), AVE-8062,BEC2 (mitumomab), cachectin or cachexin (tumor necrosis factor),canvaxin (vaccine), CeaVac™ (cancer vaccine), CELEUK® (celmoleukin),CEPLENE® (histamine dihydrochloride), CERVARIX™ (human papillomavirusvaccine), CHOP® (C: CYTOXAN® (cyclophosphamide); H: ADRIAMYCIN®(hydroxydoxorubicin); O: Vincristine (ONCOVIN®); P: prednisone), CyPat™,combrestatin A4P, DAB(389)EGF or TransMID-107R™ (diphtheria toxins),dacarbazine, dactinomycin, 5,6-dimethylxanthenone-4-acetic acid (DMXAA),eniluracil, EVIZON™ (squalamine lactate), DIMERICINE® (T4N5 liposomelotion), discodermolide, DX-8951f (exatecan mesylate), enzastaurin,EPO906, GARDASIL® (quadrivalent human papillomavirus (Types 6, 11, 16,18) recombinant vaccine), gastrimmune, genasense, GMK (gangliosideconjugate vaccine), GVAX® (prostate cancer vaccine), halofuginone,histerelin, hydroxycarbamide, ibandronic acid, IGN-101, IL-13-PE38,IL-13-PE38QQR (cintredekin besudotox), IL-13-pseudomonas exotoxin,interferon-α, interferon-γ, JUNOVAN™ or MEPACT™ (mifamurtide),lonafarnib, 5,10-methylenetetrahydrofolate, miltefosine(hexadecylphosphocholine), NEOVASTAT® (AE-941), NEUTREXIN® (trimetrexateglucuronate), NIPENT® (pentostatin), ONCONASE® (a ribonuclease enzyme),ONCOPHAGE® (melanoma vaccine treatment), OncoVAX (IL-2 Vaccine),ORATHECIN™ (rubitecan), OSIDEM® (antibody-based cell drug), OvaRex® MAb(murine monoclonal antibody), paditaxel, PANDIMEX™ (aglycone saponinsfrom ginseng comprising 20(S)protopanaxadiol (aPPD) and20(S)protopanaxatriol (aPPT)), panitumumab, PANVAC®-VF (investigationalcancer vaccine), pegaspargase, PEG Interferon A, phenoxodiol,procarbazine, rebimastat, REMOVAB® (catumaxomab), REVLIMID®(lenalidomide), RSR13 (efaproxiral), SOMATULINE® LA (lanreotide),SORIATANE® (acitretin), staurosporine (Streptomyces staurospores),talabostat (PT100), TARGRETIN® (bexarotene), Taxoprexin®(DHA-paclitaxel), TELCYTA™ (TLK286), temilifene, TEMODAR®(temozolomide), tesmilifene, thalidomide, THERATOPE® (STn-KLH), thymitaq(2-amino-3,4-dihydro-6-methyl-4-oxo-5-(4-pyridylthio)quinazolinedihydrochloride), TNFerade™ (adenovector: DNA carrier containing thegene for tumor necrosis factor-α), TRACLEER® or ZAVESCA® (bosentan),tretinoin (Retin-A), tetrandrine, TRISENOX® (arsenic trioxide),VIRULIZIN®, ukrain (derivative of alkaloids from the greater celandineplant), vitaxin (anti-alphavbeta3 antibody), XCYTRIN® (motexafingadolinium), XINLAY™ (atrasentan), XYOTAX™ (paclitaxel poliglumex),YONDELIS™ (trabectedin), ZD-6126, ZINECARD® (dexrazoxane), zometa(zolendronic acid), zorubicin and the like.

It is also expected that ABT-888 Crystalline Form 1 would inhibit growthof cells derived from a pediatric cancer or neoplasm including embryonalrhabdomyosarcoma, pediatric acute lymphoblastic leukemia, pediatricacute myelogenous leukemia, pediatric alveolar rhabdomyosarcoma,pediatric anaplastic ependymoma, pediatric anaplastic large celllymphoma, pediatric anaplastic medulloblastoma, pediatric atypicalteratoid/rhabdoid tumor of the central nervous system, pediatricbiphenotypic acute leukemia, pediatric Burkitts lymphoma, pediatriccancers of Ewing's family of tumors such as primitive neuroectodermalrumors, pediatric diffuse anaplastic Wilm's tumor, pediatric favorablehistology Wilm's tumor, pediatric glioblastoma, pediatricmedulloblastoma, pediatric neuroblastoma, pediatricneuroblastoma-derived myelocytomatosis, pediatric pre-B-cell cancers(such as leukemia), pediatric psteosarcoma, pediatric rhabdoid kidneytumor, pediatric rhabdomyosarcoma, and pediatric T-cell cancers such aslymphoma and skin cancer and the like.

The following examples are presented to provide what is believed to bethe most useful and readily understood description of procedures andconceptual aspects of this invention.

EXAMPLE 1 Preparation of ABT-888 Crystalline Form 1

A mixture of ABT-888 dihydrochloride (10 g) was stirred in saturatedpotassium bicarbonate (50 mL) and n-butanol (50 mL) until the ABT-888dihydrochloride completely dissolved. The aqueous layer was extractedwith a second portion of n-butanol then discarded. The extracts werecombined, washed with 15% sodium chloride solution (50 mL) andconcentrated. The concentrate was chase distilled three times withheptane (50 mL),dissolved in refluxing 2-propanol (45 mL) and filteredhot. The filtrate was cooled to ambient temperature with stirring over18 hours, cooled to 0-5° C., stirred for 1 hour, and filtered. Thefiltrant was washed with 2-propanol and dried in a vacuum oven at 45-50°C. with a slight nitrogen purge.

EXAMPLE 2 Preparation of ABT-888 Crystalline Form 2

A mixture of ABT-888 in methanol, in which the ABT-888 was completelydissolved, was concentrated at about 35 ° C., and the concentrate wasdried to a constant weight.

EXAMPLE 3 Preparation of ABT-888 Crystalline Form 1

-   Step 1: 2-(2-methyl-2-pyrrolidino)-benzimidazole-4-carboxamide 2    HCl (15) is dissolved in water (3.5 kg/kg 15) at 20±5° C.    Dissolution of 15 in water results in a solution of pH 0-1.-   Step 2: The reaction is run at 20-25° C. One equivalent of sodium    hydroxide is added, raising the pH to 2-3 with only a mild exotherm    (10° C. observed with rapid addition of 1.0 equiv.). This generates    a solution that remains clear for several days even when seeded with    free base crystals. 3N NaOH (1.0 equiv., 1.25 kg/kg 15) is charged    and the solution polish filtered into the crystallizer/reactor.-   Step 3: 5% Na₂CO₃ (1.5 equiv., 10.08 kg/kg 15) is then filtered into    the crystallizer over 2 hours. Nucleation occurs after approximately    ⅙th of the Na₂CO₃ solution is added (˜0.25 equiv.)-   Step 4: The slurry is mixed for NLT 15 min before sampling    (typically 1 to 4 hours (2.5 mg/mL product in the supernatant)). The    slurry is filtered at 20° C. and washed with 6 portions of water    (1.0 kg/kg 15 each). Each wash was applied to the top of the cake    and then pressured through. No mixing of the wetcake was done.-   Step 5: The solids are then dried. Drying was performed at 50° C.    keeping the Cogeim under vacuum while applying a slight nitrogen    bleed. The agitator blade was left in the cake to improve heat    transfer to the cake. It was rotated and lifted out of the cake once    per hour of drying to speed the drying process while minimizing    potential crystal attrition that occurs with continuous agitator    use.

In one embodiment of Step 1, the volume of water for dissolution of theDihydrochloride (15) is about 1.3 g water/g 15. In another embodiment ofStep 1, the volume of water for dissolution is about 1.3 g to about 4 gwater/g 15. In another embodiment of Step 1, the volume of water fordissolution is 1.3 g to 3.5 g water/g 15. In another embodiment of Step1, the volume of water for dissolution is 3.5 g water/g 15.

In one embodiment of Step 2, the pH after addition of NaOH solution isbetween about pH 11 and pH 10. In another embodiment, the pH is justunder 10. In another embodiment of Step 2, equential additions of NaOHand Na₂CO₃ were performed for neutralization. Sodium carbonate has theproperty of buffering the system in the range of 9.4 to 9.6 once 2.0equivalents (total) of base have been added (assuming a stoichiometricconversion of Na₂CO₃ to NaHCO₃). In another embodiment of Step 2, 6 NNaOH solution (21%) is used in the conversion. In another embodiment ofStep 2, Na₂CO₃ solutions as concentrated as 15% are used in theconversion.

In one embodiment of Step 3, 1.1 equivalent of base is added (1.0 equiv.NaOH, and 0.1 equiv. Na₂CO₃). In another embodiment of Step 3, fromabout 1.1 to about 1.34 equivalent of base is added. In anotherembodiment of Step 3, more than 1.34 equivalent of base is added. Inanother embodiment, 2.5 equivalent of base (1.0 equiv. NaOH plus 1.5equiv. Na₂CO₃) is added.

In another embodiment of Step 4, 3 portions of 1 kg water/kg 15 are usedin the wash. In another embodiment of Step 4, the wash is mixed with thecake to increase the contact time.

In another embodiment, crystal size and strength may be affected byinitiating crystallization through seeding prior to addition of thefirst 0.1 equivalent of sodium carbonate. In another embodiment, thebatch is cooled prior to crystallization. In another embodiment, amiscible solvent, such as ethanol or isopropanol, is added to thereaction.

Powder X-ray diffraction was performed using an XDS-2000/X-raydiffractometer equipped with a 2 kW normal focus X-ray tube and aPeltier cooled germanium solid-state detector (Scintag Inc., Sunnyvale,Calif.). The data were processed using DMSNT software (version 1.37).The X-ray source was a copper filament (Cu—Kα at 1.54178 Å) operated at45 kV and 40 mA. The alignment of the goniometer was checked daily usinga Corundum standard. The sample was placed in a thin layer (with noprior grinding) onto a zero background plate and continuously scanned ata rate of 2° 2θ per minute over a range of 2°-40° 2θ.

It is meant to be understood that relative intensities of peak heightsin a PXRD pattern may vary and will be dependent on variables such asthe temperature, size of crystal size or morphology, sample preparation,or sample height in the analysis well of the X-ray diffractometer.

It is also meant to be understood that peak positions may vary whenmeasured with different radiation sources. For example, Cu—Kα₁, Mo—Kα,Co—Kα and Fe—Kα radiation, having wavelengths of 1.54060 Å, 0.7107 Å,1.7902 Å and 1.9373 Å, respectively, may provide peak positions whichdiffer from those measured with Cu—Kα radiation, which has a wavelengthof 1.5478 Å.

The term “about” preceding a series of peak positions means that all ofthe peaks of the group which it precedes are reported in terms ofangular positions (two theta) with an allowable variability of ±0.1° asspecified by the U.S. Pharmacopeia, pages 1843-1884 (1995). Thevariability of ±0.1° is intended to be used when comparing two powderX-ray diffraction patterns. In practice, if a diffraction pattern peakfrom one pattern is assigned a range of angular positions (two theta)which is the measured peak position ±0.1° and if those ranges of peakpositions overlap, then the two peaks are considered to have the sameangular position. For example, if a peak from one pattern is determinedto have a position of 11.0°, for comparison purposes the allowablevariability allows the peak to be assigned a position in the range of10.9°-11.1°.

Accordingly, for example, the phrase “about 9.9°, 11.0° and 11.8° andone or more than one additional peaks having respective 2θ values ofabout 14.6°, 15.2°, 18.2°, 19.6°, 20.3°, 21.3°, 22.5°, 22.8°, 24.7°,28.5° and 29.1°,” as used herein, means about 9.9°, about 11.0° andabout 11.8° and one or more than one additional peaks having respective2θ values of about 14.6°, about 15.2°, about 18.2°, about 19.6°, about20.3°, about 21.3°, about 22.5°, about 22.8°, about 24.7°, about 28.5°and about 29.1° and also means 9.9°±0.1°, 11.0°±0.1° and 11.8°±0.1° andone or more than one additional peaks having respective 2θ values ofabout 14.6°±0.1°, 15.2°±0.1°, 18.2°±0.1°, 19.6°±0.1°, 20.3°±0.1°,21.3°±0.1°, 22.5°±0.1°, 22.8°±0.1°, 24.7°±0.1°, 28.5°±0.1° and29.1°±0.1°.

The term “about” preceding a temperature means the given temperature ±2°C. For example, about 25° C. means 25° C.±2° C. or 23° C.-27° C.

Heat flow was measured using a differential scanning calorimeter (model2920 with Thermal Advantage version 1. 1A operating software (TAInstruments, New Castle, Del.). A sample (1-4 mg) was weighed into analuminum pan, and the pan was covered with and aluminum lid containing apinhole to allow vapor to escape. The partially sealed pan was placed inthe furnace and heated in an open pan at a rate of 10° C./min. Indiumstandards were used for temperature and heat of fusion calibration. Dataanalysis was performed using separate software (Universal Analysis forWindows 2000/XP, version 4.2E, TA Instruments, New Castle, Del.).

As shown in FIG. 3, when heated at 10° C./min, ABT-888 Crystalline Form1 exhibits a single endothermic event, corresponding to melting at188.3° C. Multiple determinations showed a melting point 188.6±0.8° C.

Raman spectra were measured using a laser Raman spectrometer (RamanRxn1HTS-785 785 nm NIR laser and model HLS-L microprobe, with Holograms™version 4.004.0.0230 and Holomap™ version 2.3.4 software, Kaiser OpticalSystems, Inc., Ann Arbor, Mich.).

Transmission infrared spectra of the solids were obtained using aFourier-transform infrared spectrometer (Nicolet Magna 750 FT-IRSpectrometer, Nicolet Instrument Corporation, Madison, Wis.) equippedwith a Nicolet NIC-PLAN Microscope and MCT-A liquid nitrogen cooleddetector. The sample was placed on a 13 mm×1 mm BaF₂ disc sample holderand 64 scans were collected at 4 cm⁻¹ resolution.

The foregoing is meant to be illustrative of the invention and notintended to limit it to the disclosed embodiments. Variations andchanges obvious to one skilled in the art are intended to be within thescope and nature of the invention as defined in the claims.

1. 2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1 which, when measured at about −100° C. in atetragonal crystal system and P4₃2₁2 space group with radiation at0.7107 Å, is characterized by respective lattice parameter values a, band c of 8.218 Å±0.002 Å, 8.218 Å±0.002 Å and 36.06 Å±0.01 Å and α, βand γ values of 90°. 2.2-((R)-2-Methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamideCrystalline Form 1 which, when measured at about 25° C. with radiationat 1.54178 Å, is characterized by a powder diffraction pattern havingrespective 2θ values of about 9.9°, 11.0° and 11.8° and one or more thanone additional peaks having respective 2θ values of about 14.6°, 15.2°,18.2°, 19.6°, 20.3°, 21.3°, 22.5°, 22.8°, 24.7°, 28.5° and 29.1°.